With the development of DNA sequencing technologies, high-throughput sequencing technologies have been widely applied in various fields of life science research. Although the cost of sequencing technologies is becoming lower and lower with the on-going updating and popularization of sequencing technologies, the expense of whole-genome sequencing technologies per se is still costly. A preferable solution to this problem would be to enrich for a target region of interest before conducting high-throughput sequencing. Conventional sequence capture techniques typically involve constructing a high-throughput sequencing library, then enriching the library for a target region using a probe prior to conducting sequencing.
Additionally, Life Tech Corporation developed a kit called Ampliseq based on multiplex PCR technology. The kit allows to achieve enrichment of a target region through a multiplex PCR process, thus greatly shortening the time-consuming operation process of capture by hybridization. This represents an innovation in region capture technology.
However, Ampliseq is unable to enrich for a target region in a cellular free DNA, because the cellular free DNA is characterized by small fragment length, which may even be smaller than the length of the PCR products in the Ampliseq kit. Moreover, the region capture technology based on Ampliseq can achieve pooling of samples only after performing PCR, which would be a time-consuming process.